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. 2008 Jul 28;36(15):5074–5082. doi: 10.1093/nar/gkn489

Figure 2.

Figure 2.

Subunit θ solubilizes nascent ɛ and α:ɛ:θ forms a soluble isolable complex when ɛ is made in the presence of α and θ. The ɛ-subunit was produced by cell-free synthesis. The gels were stained with Coomassie brilliant blue. T, P and S denote the total reaction mixtures, the insoluble fractions (pellet) and the supernatants, respectively. (A) The 10% SDS–PAGE of ɛ synthesized at 37°C in the presence of 5 mg/ml of α-subunit. Nascent ɛ coprecipitates α. (B) The 15% SDS–PAGE of ɛ synthesized at 37°C in the presence of 0.1 mg/ml (lanes 1–3, labeled ɛ) or 1 mg/ml (lanes 4–6, labeled ɛ + θ) of θ-subunit. (C) The 15% SDS–PAGE of ɛ synthesized at 30°C in the presence of 5 mg/ml of the α-subunit and 0.5 mg/ml of the θ-subunit. The left and right lanes show the soluble fraction of the cell-free reaction mixture and the purified α:ɛ:θ complex, respectively.